SUBUNITS OF HUMAN CHORIONIC GONADOTROPHIN: AN IMMUNOCHEMICAL STUDY

1973 ◽  
Vol 73 (1) ◽  
pp. 133-145 ◽  
Author(s):  
S. Donini ◽  
V. Olivieri ◽  
G. Ricci ◽  
P. Donini
Keyword(s):  
Gel Filtration ◽  
Cross Reactivity ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Physical Characterization ◽  
Human Pituitary ◽  
Immunochemical Study ◽  
Urea Treatment ◽  
Α And Β Subunits

ABSTRACT The α and β subunits of HCG were prepared by urea-treatment of the hormone and isolated by DEAE-Sephadex A-25 chromatography. Further purification of the subunits was achieved by gel filtration on Sephadex G-100. Partial chemico-physical characterization confirmed the non-identity of the HCG subunits, as previously reported by several investigators. A radioimmunoassay (RIA) system based on 125I-HCG-β + anti-HCG-β serum was used to study the antigenic similarities between HCG, HCG-α and -β, human pituitary LH, urinary LH and FSH. The highest cross-reactivity was found for HCG, followed by HCG-α, LH and FSH. The specificity of the above mentioned RIA system was increased by absorbing the anti-HCG-β serum with HCG-α. Because of the ability of this absorbed antiserum to discriminate between HCG and LH, a study was carried out to detect specifically HCG in urine in spite of the presence of physiological amounts of LH.

SUBUNITS OF HUMAN CHORIONIC GONADOTROPHIN: IMMUNOLOGICAL AND BIOLOGICAL STUDIES

1975 ◽  
Vol 79 (4) ◽  
pp. 749-766 ◽  
Author(s):  
S. Donini ◽  
I. D'Alessio ◽  
P. Donini
Keyword(s):  
Biological Activity ◽  
Gel Filtration ◽  
Cross Reactivity ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Biologically Active ◽  
Β Subunit ◽  
Reactive Material ◽  
Biological Studies ◽  
The Cross

ABSTRACT The α and β subunits of human chorionic gonadotrophin (hCG) were prepared by incubation in 8 m urea, pH 4.5. The separation of the two subunits was obtained by DEAE-Sephadex A-25 chromatography and purification was carried out by gel filtration on Sephadex G-100. The β subunit obtained was biologically active and was therefore further purified by affinity chromatography using as immuno-adsorbent the α antibodies coupled to Sepharose 4B. The β subunit so purified showed a biological activity less than 1 IU/mg. The immunological and biological properties of the hCG subunits have been studied. It was found that the anti hCG β serum can discriminate between hCG and hLH and that in the 125I-hCG + anti-β serum radioimmunoassay, the cross-reactivity of pituitary hLH was lower than that of urinary hLH. Moreover, it was observed that the less purified was the urinary LH preparation, the higher was the cross-reactivity. Therefore we considered the hypothesis that during the purification of human menopausal gonadotrophin (hMG) some LH subunits or smaller immunoreactive fragments could have been discarded with the waste fractions. In order to test the validity of this hypophysis, all the protein fractions obtained during the purification of the hMG were gel-filtered on Sephadex G-100. The immunoreactivity of the effluents from the gel filtration was tested by hCG, hCG-β, hCG-α and hLH radioimmunoassays. While the α reactive material was found in some fractions as a peak having the same Ve/Vo value as hCG-α, the β reactive material present in the crude hMG fractions was not observed in other fractions. The cross-reactivity with the anti β serum was very low and was found in the LH region of the gel chromatogram. Furthermore, the neutralization of the biological activity of hCG and of urinary and pituitary LH by the anti hCG β serum was studied by incubating a fixed amount of the three hormones with increasing volumes of antiserum and measuring the LH activity after incubation by the OAAD test. It was observed that the anti hCG β serum inhibits hCG more than urinary or pituitary LH.

Studies of the human chorionic gonadotrophin-like substance of human pituitary glands and its significance

1983 ◽  
Vol 96 (1) ◽  
pp. 115-126 ◽  
Author(s):  
Anne Stockell Hartree ◽  
Ratna C. Shownkeen ◽  
V. C. Stevens ◽  
Shuji Matsuura ◽  
Masanobu Ohashi ◽  
...  
Keyword(s):  
Gel Filtration ◽  
Molecular Size ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Partial Purification ◽  
Human Pituitary ◽  
Human Pituitary Gland ◽  
Diethylaminoethyl Cellulose ◽  
Pituitary Glands ◽  
Pregnancy Urine

Radioimmunoassays utilizing antisera specific for the carboxyl-terminal portion of human chorionic gonadotrophin (hCG) β-subunit were used to measure the concentration in human pituitary extracts of an immunoactive hCG factor (hCG′) which was different from human LH (hLH). The content of hCG′ from different human pituitary pools collected between 1966 and 1979 was relatively constant at 0·5–1·1 μg per gland. The hCG′ concentrations observed in acetone-dried powder of individual human pituitary glands (0·4–26 ng/mg) were close to those reported for full-term placental powder. After separation and partial purification of human pituitary glycoprotein hormones, pituitary hCG′ was found mainly in the crude human FSH (hFSH) fraction. It was separated from hFSH by diethylaminoethyl–cellulose chromatography at pH 7 and by gel filtration on Sephadex G-100. On gel filtration its molecular size was larger than that of hLH or hFSH and it was strongly bound to Concanavalin A–Sepharose. The most active preparations of pituitary hCG' obtained by these procedures were approximately 5 per cent as potent by specific radioimmunoassay as hCG purified from pregnancy urine. Although the hCG′ content in individual pituitary glands was more variable than the hLH content, on average pituitary hCG′ was estimated to be around 25- to 50-fold less than the content of hLH, hFSH or human TSH in the human pituitary gland.

Therapeutic use of gonadotrophins and clomiphene

1969 ◽  
Vol 7 (9) ◽  
pp. 33-35
Keyword(s):  
Pregnant Women ◽  
Follicle Stimulating Hormone ◽  
Therapeutic Use ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Human Pituitary ◽  
Synthetic Compound ◽  
Pituitary Glands ◽  
Post Menopausal ◽  
Stimulating Hormone

The three substances now used to stimulate the gonads in infertility are human follicle stimulating hormone (HFSH) obtained mainly from post-menopausal urine, but also from human pituitary glands, human chorionic gonadotrophin (HCG) extracted from the urine of pregnant women, and clomiphene (Clomid - Merrell), a synthetic compound which we reviewed in 1967.1

STUDIES ON A HUMAN CHORIONIC GONADOTROPHIN-LIKE MATERIAL PRESENT IN NON-PREGNANT SUBJECTS

1978 ◽  
Vol 89 (3) ◽  
pp. 492-505 ◽  
Author(s):  
D. M. Robertson ◽  
H. Suginami ◽  
H. Hernandez Montes ◽  
C. P. Puri ◽  
S. K. Choi ◽  
...  
Keyword(s):  
Gel Filtration ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Specific Sequence ◽  
Gonadotrophin Secretion ◽  
Carboxy Terminal Region ◽  
Assay Methods ◽  
Low Levels ◽  
Carboxy Terminal

ABSTRACT The presence of an hCG-like material in urinary and pituitary extracts and plasma obtained from non-pregnant subjects was investigated. Two assay methods were used to detect this material following fractionation of pituitary and urinary extracts by gel filtration (Ultrogel AcA 54) and/or isoelectrofocusing: a) a radioimmunoassay employing an antiserum raised against a specific sequence of the carboxy terminal region (residues 115– 145) of the β-subunit of hCG, and b) an in vitro bioassay method which measures both hLH and hCG activities. The fractionation procedures employed provide a satisfactory separation of highly purified hCG and hLH preparations. In the pituitary and urinary extracts hCGβ-peptide-like immunoactive (PIA) material was found consistently, which co-eluted with iodinated hCG following gel filtration and possessed pI values similar to those of hCG when subjected to isoelectrofocusing. The PIA material also exhibited in vitro biological activity similar to that shown by hLH and hCG. Detectable levels of immunoactive material were also found in plasma; however, the plasma levels of this PIA material were not influenced by classical endocrine measures such as the stimulation or inhibition of gonadotrophin secretion. The low levels of this material in plasma precluded its further characterization by gel filtration or electrofocusing. Whereas the present data and those reported by other investigators seem to suggest the presence of some hCG-like material in urinary and pituitary extracts and possibly in plasma of non-pregnant subjects, it is emphasized that the available evidence is not sufficiently conclusive to exclude other interpretations as to the nature of this material.

OBSERVATIONS ON THE ASSAY OF HUMAN URINARY FOLLICLE-STIMULATING HORMONE BY THE AUGMENTATION TEST IN MICE

1966 ◽  
Vol 35 (2) ◽  
pp. 199-206 ◽  
Author(s):  
P. S. BROWN ◽  
M. WELLS
Keyword(s):  
No Value ◽  
Follicle Stimulating Hormone ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
High Potency ◽  
Human Pituitary ◽  
Ovarian Weight ◽  
C3h Mice ◽  
C57bl Mice ◽  
Stimulating Hormone

SUMMARY The follicle-stimulating hormone (FSH) content of urinary gonadotrophic extracts was assayed by its effect on the ovarian weight of immature mice when given in conjunction with 40 i.u. human chorionic gonadotrophin. About three-quarters of all routine assays gave values of λ between 0·15 and 0·30. Precision was slightly increased when the material was given in three rather than in five injections. Correction of ovarian weight for body weight was either invalid or of no value in reducing variance. Removal of between-litter variance increased precision considerably. Mice of three randomly bred colonies were all satisfactory, and inbred C57BL mice were also suitable for the assay. C3H mice were less sensitive. The efficiency of different methods of extracting FSH from urine was examined. The method of Johnsen (1958) using precipitation with tannic acid was considered the most satisfactory and gave extracts of high potency and low bulk. Limited experiments in which purified human pituitary FSH was assayed with and without added luteinizing hormone, gave results compatible with the assumption that the method is specific for FSH.

IMMUNOLOGICAL DETERMINATION OF PITUITARY LUTEINIZING HORMONE IN THE URINE OF FERTILE AND POST-MENOPAUSAL WOMEN AND ADULT MEN

1962 ◽  
Vol 39 (4) ◽  
pp. 539-546 ◽  
Author(s):  
Leif Wide ◽  
Carl Gemzell
Keyword(s):  
Luteinizing Hormone ◽  
Haemagglutination Inhibition ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Human Pituitary ◽  
Adult Men ◽  
Menopausal Women ◽  
Post Menopausal ◽  
Inhibition Reaction

ABSTRACT An immunological method to assay human pituitary luteinizing hormone (HPLH) in urine is described. It is based on the fact that HPLH crossreacts with human chorionic gonadotrophin (HCG) in an haemagglutination inhibition reaction between HCG-coated blood cells and rabbit HCG-antisera. During the menstrual cycle the excretion of HPLH reached a peak of 200–400 U per liter at the time of ovulation. In the urine of post-menopausal women the concentration of HPLH was between 100 and 400 U per liter. In the urine of adult men the concentration of HPLH was between 50 and 160 U per liter.

INTERNATIONAL REFERENCE PREPARATION OF HUMAN CHORIONIC GONADOTROPHIN FOR IMMUNOASSAY: POTENCY ESTIMATES IN VARIOUS BIOASSAY AND PROTEIN BINDING ASSAY SYSTEMS; AND INTERNATIONAL REFERENCE PREPARATIONS OF THE α AND β SUBUNITS OF HUMAN CHORIONIC GONADOTROPHIN FOR IMMUNOASSAY

1980 ◽  
Vol 84 (2) ◽  
pp. 295-310 ◽  
Author(s):  
P. L. STORRING ◽  
ROSE E. GAINES-DAS ◽  
D. R. BANGHAM
Keyword(s):  
Geometric Mean ◽  
Collaborative Study ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Considerable Heterogeneity ◽  
International Reference ◽  
Reference Preparation ◽  
Α And Β Subunits

The preparation and nature of the International Reference Preparation of Human Chorionic Gonadotrophin (HCG) for Immunoassay (IRP), as well as that of a second batch of ampoules (HCG 75/589) prepared identically from the same HCG preparation, are described. A collaborative study of these materials was carried out by 11 laboratories in eight countries, using different bioassay and immunoassay methods. Using the various in-vivo and in-vitro bioassays and receptor assays, the mean log potency estimates for each method within each laboratory of the HCG content of ampoules of the IRP, in terms of the Second International Standard of Human Chorionic Gonadotrophin for Bioassay (IS), were homogeneous and gave an overall weighted geometric mean (95% confidence limits) of 650 (632–669) International Units (i.u.)/ampoule. There was considerable heterogeneity of potency estimates of the IRP in terms of the IS both within and between many of the immunoassay systems (reflecting the impurity of the IS), and hence attempts to calibrate the IRP with immunoassay systems of different specificities were invalid. Immunoassay estimates of the HCG content of preparations of serum and urine, in terms of the IRP, showed considerable heterogeneity between assay systems (although the degree of this heterogeneity was no greater than that observed using the IS as standard), but the ranking order between preparations was consistent. Confirmation was obtained that contamination of the IRP with HCG-α and HCG-β subunits was insignificant. Accelerated degradation studies of the IRP stored at increased temperatures suggested that its stability under normal storage conditions would be satisfactory. It was agreed that the IRP was suitable to serve as an international reference preparation for immunoassay, and it was assigned a unitage of 650 i.u./ampoule on the basis of bioassay calibration. Since the ampoules of HCG (75/589) did not differ significantly from the IRP in any of the assay systems studied, it appeared to be equally suitable as a reference preparation. The International Reference Preparations of the α and β Subunits of Human Chorionic Gonadotrophin for Immunoassay are also described.

scholarly journals Molar pregnancy and thyroid storm - literature review

2017 ◽  
Vol 23 (3) ◽  
pp. 121-125 ◽  
Author(s):  
G. A. Filipescu ◽  
Oana Alina Solomon ◽  
Nicoleta Clim ◽  
Amelia Milulescu ◽  
Andreea Gratiana Boiangiu ◽  
...  
Keyword(s):  
Luteinizing Hormone ◽  
Literature Review ◽  
Follicle Stimulating Hormone ◽  
Cross Reactivity ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Thyroid Storm ◽  
Unusual Complication ◽  
Molar Pregnancy ◽  
Dilation And Curettage

AbstractMolar pregnancies results from a tainted fertilization process. Trophoblastic thyroidian hyper function is an unusual complication of a molar pregnancy. The degree of thyroid stimulation and the severity of clinical hyperthyroidism is directly proportional to HCG concentration. Human chorionic gonadotrophin is almost identical with TSH, luteinizing hormone (LH) and follicle-stimulating hormone, this analogy in the structure will cause cross-reactivity with their receptors. Hyperthyroid status can vary from asymptomatic hyper function to thyroid storm. Dilation and curettage represents the treatment for hyperthyroidism in molar pregnancy. Awareness of this condition is important for diagnosis and treatment.

RELATIONSHIPS BETWEEN PHYSICO-CHEMICAL IMMUNOLOGICAL AND BIOLOGICAL PROPERTIES OF HUMAN CHORIONIC GONADOTROPHIN

1971 ◽  
Vol 67 (3) ◽  
pp. 445-456 ◽  
Author(s):  
D. L. Matthies ◽  
P. Petrusz ◽  
E. Diczfalusy
Keyword(s):  
Follicle Stimulating Hormone ◽  
Gel Filtration ◽  
Biological Properties ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Gel Filtration Chromatography ◽  
Physico Chemical ◽  
Immunological Test ◽  
Stimulating Hormone

ABSTRACT Purified human chorionic gonadotrophin (HCG) preparations were chromatographed on Sephadex G-100 and the distribution of the hormone on the chromatogram was studied by an immunological test and by bioassay. Each preparation studied showed evidence of inhomogeneity in terms of the degree to which it was retarded on the gel bed, but the type of inhomogeneity was variable. By the use of gel filtration chromatography it was possible to separate HCG molecules devoid of detectable follicle stimulating hormone (FSH)-like activity from molecules which possessed FSH-like activity. Evidence is presented which indicates that at least some physico-chemical manipulations utilized in the purification of HCG from urine may introduce artifacts.

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